Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From September 26, 2016 to January 2, 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
did not affect outcome/integrity of the study
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Deviations:
yes
Remarks:
did not affect outcome/integrity of the study
Qualifier:
according to guideline
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Deviations:
yes
Remarks:
did not affect outcome/integrity of the study
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

Constituent 1
Reference substance name:
Hexane, 1,6-diisocyanato-, homopolymer, 2-hydroxyethyl acrylate- and propylene glycol monoacrylate-blocked
EC Number:
806-984-5
Cas Number:
1392411-89-0
IUPAC Name:
Hexane, 1,6-diisocyanato-, homopolymer, 2-hydroxyethyl acrylate- and propylene glycol monoacrylate-blocked
Test material form:
liquid: viscous
Details on test material:
Name: Ebecryl 4738, Batch no.: DR0004393

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Crl:WI(Han)
Details on species / strain selection:
Test system Rat: Crl:WI(Han) (outbred, SPF-Quality).
Source: Charles River Deutschland, Sulzfeld, Germany.
Nulliparous and nonpregnant females and untreated animals were used at initiation of the study.
Rationale: This species and strain of rat has been recognized as appropriate for general and reproduction toxicity studies Charles River Den Bosch has general and reproduction/developmental historical data in this species from the same strain and source.
Sex:
male/female
Details on test animals or test system and environmental conditions:
Age: Approximately 10-12 weeks;
Number of animals: 40 females and 40 males;
Acclimatization: At least 5 d prior to start of treatment;
Health inspection: At least upon receipt of the animals;
Randomization: Before initiation of treatment, by computer-generated random algorithm according to body weight, with all animals within ± 20% of the sex mean;
Identification: Earmark and tattoo.

Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, at least 10 air changes/h, and a 12-h light/12-h dark cycle. The light/dark cycle was interrupted for study related activities.

Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF Spezialdiäten GmbH, Soest, Germany). During motor activity measurements, animals did not have access to food for a maximum of approximately 2.5 h. There has been provided a free access to tap-water.

Administration / exposure

Route of administration:
oral: gavage
Details on route of administration:
Oral gavage, using a plastic feeding tube. Formulations were placed on a magnetic stirrer during dosing.
Vehicle:
propylene glycol
Details on oral exposure:
Based on the results of a dose range finding study in which dose levels of 500 and 1000 mg/kg bw were tested (17 and 10 d, respectively), no dose limiting effects were seen, the dose levels for the main study were therefore selected to be 100, 300 and 1000 mg/kg bw/d.
Analytical verification of doses or concentrations:
yes
Remarks:
LC-MS/MS
Details on analytical verification of doses or concentrations:
No test substance was detected in the control group formulation. The concentrations analyzed in the formulations of groups 2, 3 and 4 (100, 300 and 1000 mg/kg bw/d) were in agreement with the target concentrations (i.e. mean accuracies between 90% and 110%). The formulations of Group 2 and Group 4 prepared were homogeneous (i.e. coefficient of variation ≤ 10%).
Formulations at the entire range were stable when stored at room temperature protected from light for at least 5 h (i.e. relative difference ≤ 10%).
The test substance was stable in propylene glycol over a storage period of at least 86 d at a temperature ≤ -70°C.
A dose control system was used as additional check to verify the dosing procedure according to Standard Operating Procedures.
Duration of treatment / exposure:
- Males were dosed for 29 d i.e. 2 weeks prior to mating, during mating, and up to the day prior to scheduled necropsy.
- Females that delivered were exposed for 40-45 d i.e. 2 weeks prior to mating (with the objective of covering at least two complete estrous cycles), the variable time to conception, the duration of the pregnancy and at least 4 d after delivery up to and including the day before scheduled necropsy. Females which failed to deliver healthy offspring were exposed for 40 or 42 d.
Frequency of treatment:
Once daily for 7 d per week
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Remarks:
Group 1
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Remarks:
Group 2
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Remarks:
Group 3
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
Remarks:
Group 4
No. of animals per sex per dose:
10
Control animals:
yes, concurrent no treatment
Details on study design:
The test substance, formulated in propylene glycol, was administered daily by oral gavage to SPF-bred Wistar Han rats. One control group and three treated groups (100, 300 and 1000 mg/kg bw/day) of the test substance were tested, each consisting of 10 males and 10 females.
Positive control:
No positive control was used.

Examinations

Observations and examinations performed and frequency:
Mortality/Viability: At least twice daily.

Clinical signs: At least once daily from start of treatment up to the day prior to necropsy, detailed clinical observations were made for all animals, at least 1 h (± 30 min) after dosing (based on the peak period of anticipated effects after treatment). Once prior to start of treatment and at weekly intervals during the treatment period this was also performed outside the home cage in a standard arena. The time of onset, grade and duration of any observed sign was recorded. Signs were graded for severity and the maximum grade was predefined at 3 or 4. Grades were coded as slight (grade 1), moderate (grade 2), severe (grade 3) and very severe (grade 4). For certain signs, only its presence (grade 1) or absence (grade 0) was scored. In the data tables, the scored grades were reported, as well as the percentage of animals affected in summary tables.

Functional Observations: The following functional observations tests were performed on each individual animal of the selected 5 animals/sex/group:
• hearing ability (HEARING), pupillary reflex (PUPIL L/R), and static righting reflex (STATIC R) (Score 0 = normal/present, score 1 = abnormal/absent).
• fore- and hind-limb grip strength, recorded as the mean of three measurements per animal.
• locomotor activity (recording period: 1-hour under normal laboratory light conditions, using a computerized monitoring system.
Total movements and ambulations are reported. Ambulations represent movements characterized by a relocation of the entire body position like walking, whereas total movements represent all movements made by the animals, including ambulations but also smaller or finer movements like grooming, weaving or movements of the head.
The selected males were tested during Week 4 of treatment and the selected females were tested during the lactation period from Day 4 of lactation onwards (all before blood sampling). These tests were performed after observation for clinical signs (incl. arena observation, if applicable).

Body weights: Males and females were weighed on the first day of exposure (prior to first exposure) and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on PND 1 and 4.

Food consumption: Weekly, except for males and females which were housed together for mating and for females without evidence of mating. Food consumption of mated females was measured on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on PND 1 and 4.

Water consumption: Subjective appraisal was maintained during the study, but no quantitative investigation was introduced as no treatment related effect was suspected.

Haematology and clinical biochemistry parameters were evaluated. Samples collection:
Blood samples were collected at the end of the treatment period on the day of scheduled necropsy from the selected 5 animals/sex/group under anaesthesia using isoflurane between 7.00 and 10.30 a.m. The selected animals were deprived of food overnight (with a maximum of 24 hours) before blood sampling, but water was available. Blood samples were drawn from the retro-orbital sinus and collected into tubes prepared with K3-EDTA for haematological parameters (0.5 mL), with citrate for clotting tests (0.45 mL) and tubes treated with Li-heparin for clinical biochemistry parameters (0.5 mL). An additional blood sample (0.25 mL) was collected into serum tubes for determination of bile acids.
Sacrifice and pathology:
- Necropsy: 5 animals/sex/group - Males following completion of the mating period, females which delivered, females which failed to deliver (females with evidence of mating).

- Gross lesions of all animals were recorded (all dose groups).

- Terminal body weights were recorded from all males and the selected 5 females/group. The following organ weights were recorded from the following animals on the scheduled day of necropsy: adrenal glands, brain, epididymides, heart, kidneys, liver, ovaries, prostate, seminal vesicles including coagulating glands, spleen, testes, thymus, thyroid (including parathyroid), uterus (including cervix).

- Pathology examinations:
The numbers of corpora lutea and former implantation sites were recorded for all paired females. Samples of the following tissues and organs were collected and fixed in 10% buffered formalin for further histophatological examinations: ovaries, adrenal glands, pancreas, aorta, peyer's patches, brain (cerebellum, mid-brain, cortex), pituitary gland, caecum, preputial gland, cervix, prostate gland, clitoral gland rectum, colon, salivary glands (mandibular, sublingual), coagulation gland sciatic nerve, duodenum seminal vesicles, epididymide skeletal muscle, eyes, skin, female mammary gland area, spinal cord (cervical, midthoracic, lumbar), femur including joint, spleen, heart, sternum with bone marrow, ileum, stomach, jejunum, testes, epididymides, kidneys, thymus, lacrimal gland, exorbital, thyroid including parathyroid, larynx, tongue, liver, trachea, lung, urinary bladder.




Statistics:
Statistical Analyses:
The following statistical methods were used to analyse the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel test was applied if the data could not be assumed to follow a normal distribution.
- The Fisher Exact test was applied to frequency data.
- The Kruskal-Wallis nonparametric ANOVA test was applied to motor activity data to determine intergroup differences.

Results and discussion

Results of examinations

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
- No treatment-related clinical signs of toxicity were noted during daily clinical observations or during weekly arena observations.
- Salivation occurred after dosing in all males and females treated with the test substance, generally starting in the first week of treatment. This salivation was considered to be a physiological response rather than a sign of systemic toxicity considering its slight severity and the time of occurrence (i.e. after dosing), and may be related to the irritancy of the test substance. In addition, most treated rats showed slight lethargy after dosing on the first day of the treatment period. This clinical sign was considered not to be toxicologically relevant because it was no longer observed during the remainder of the treatment period. Any other clinical signs noted incidentally occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study and showed no dose-related trend. At the incidence observed, these were considered to be unrelated to treatment.
Mortality:
no mortality observed
Description (incidence):
No mortality occurred during the study period.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Body weight gain was higher during the first two weeks of the treatment period in males and females treated at 1000 mg/kg bw/d. Thereafter, growth rate at 1000 mg/kg bw/d was comparable to that in controls (total weight gain in males remained above control levels until the end of the study). Mean body weights of 1000 mg/kg bw/d males were close to control values (about 3% higher) while 1000 mg/kg bw/d females had statistically significantly higher body weights on Days 1 and 4 of lactation (relative difference 7%). As the changes in body weight (gain) were slight, they were considered to be of no toxicological significance. Body weight and body weight gain were not affected by treatment at 100 or 300 mg/kg bw/d.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
Food consumption was generally similar between treated and control animals. The slightly higher values noted in females at 300 and/or 1000 mg/kg bw/d at the end of the post-coitum period (statistically significant for absolute values) and during lactation (not statistically significant) were consistent with the slightly higher body weights of these females.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Description (incidence and severity):
Haematological parameters were not affected by treatment. One female at 1000 mg/kg bw/d had outlying values for platelets (higher) and red blood cell parameters (lower number of red blood cells and haemoglobin, with compensatory changes consisting of higher reticulocytes, red blood cell distribution width (RDW) and meancorpuscular volume (MCV), and increased extramedullary hematopoiesis in the spleen and liver). The reduction in red blood cells and haemoglobin could be due to blood loss at parturition. As these changes were observed in only a single animal and recovery was ongoing, they were considered not to reflect adverse effects of the test substance.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
No toxicologically relevant changes were noted in clinical biochemistry parameters of treated rats. The variations noted in clinical biochemistry values, including those achieving statistical significance, were unrelated to treatment or not toxicologically relevant due to the slight magnitude of the difference from controls (values in treated rats remained within normal limits) and/or the direction of the change.
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
Test substance-related higher liver weights (absolute and/or relative to body weight) were noted at 1000 mg/kg bw/d in both sexes. Relative differences from the control group are noted below.
Males dose level / relative to body weigh liver growth:
100 mg/kg bw: 3
300 mg/kg bw: 1
1000 mg/kg bw: 8
Females dose level, relative to body weight:
100 mg/kg bw: -5
300 mg/kg bw: 2
1000 mg/kg bw: 6 (P > 0.05)
This change was regarded as non-adverse based on its small magnitude and the absence of corroborative changes indicative of hepatotoxicity.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
All of the recorded macroscopic findings were within the range of background gross observations encountered in rats of this age and strain. These necropsy findings were therefore considered to be unrelated to treatment.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
There were no treatment-related microscopic observations. All of the recorded microscopic findings were within the range of background pathology encountered in rats of this age and strain. There was no test substance-related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.
Histopathological findings: neoplastic:
not examined
Other effects:
not examined

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects

Target system / organ toxicity

Key result
Critical effects observed:
no

Applicant's summary and conclusion

Conclusions:
Under the study conditions, there were no toxicologically relevant changes in the in-life parameters up to the highest dose of 1000 mg/kg bw/d and the repeat dose NOAEL of the substance was determined to be >1000 mg/kg bw/d in rats.
Executive summary:

A study was conducted to determine the repeated dose toxicity of the substance according to OECD Guideline 422, in compliance with GLP. The test substance, formulated in propylene glycol, was administered daily by oral gavage to SPF-bred Wistar Han rats. One control group and three test substance groups (100, 300 and 1000 mg/kg bw/d) were tested, each consisting of 10 males and 10 females. Males were dosed for 29 d, i.e. 2 weeks prior to mating, during mating, and up to the day prior to scheduled necropsy. Females were exposed for 40 -45 d, i.e. 2 weeks prior to mating (with the objective of covering at least two complete estrous cycles), the variable time to conception, the duration of the pregnancy and at least 4 d after delivery up to and including the day before scheduled necropsy. The following observations and examinations were performed: mortality, clinical signs (daily), functional observations and locomotor activity (end of treatment), body weight and food consumption (at least at weekly intervals), clinical pathology (end of treatment), gross pathology (at termination), organ weights, and histopathology on a selection of tissues. A few non-adverse, treatment-related findings were observed during the in-life phase in both sexes. Males and females treated at 1000 mg/kg bw/d had statistically significant higher liver weights (absolute and/or relative to body weight). This change was regarded as non-adverse due to its small magnitude and the absence of corroborative changes indicative of hepatotoxicity. No treatment-related or toxicologically relevant changes were recorded in clinical pathology parameters, remaining organ weights, or at macroscopic and microscopic examinations. Under the study conditions, there were no toxicologically relevant changes in the treated rats up to the highest tested dose of 1000 mg/kg bw/d and the repeat dose NOAEL of the substance was determined to be >1000 mg/kg bw/d (de Raaf - Beekhuijzen, 2017).